Creating Optimal Western Blot Conditions for OPA1 Isoforms in Skeletal Muscle Cells and Tissue
OPA1 is a dynamin-related GTPase that modulates mitochondrial dynamics and cristae integrity. Humans carry eight different isoforms of OPA1 and mice carry five, all of which are expressed as short- or long-form isoforms. These isoforms contribute to OPA1's ability to control mitochondrial energetics and DNA maintenance. However, western blot isolation of all long and short isoforms of OPA1 can be difficult. To address this issue, we developed an optimized western blot protocol based on improving running time to isolate five different isoforms of OPA1 in mouse cells and tissues. This protocol can be applied to study changes in mitochondrial structure and function. © 2025 The Author(s). Current Protocols published by Wiley Periodicals LLC. Basic Protocol: Western Blot Protocol for Isolating OPA1 Isoforms in Mouse Primary Skeletal Muscle Cells.
Authors
- Antentor Hinton (ORCID: https://orcid.org/0000-0002-7730-952X)
- Andrea G. Marshall (ORCID: https://orcid.org/0000-0001-9997-4478)
- Larry Vang (ORCID: https://orcid.org/0000-0001-9098-3650)
- Neng Vue
- Edgar Garza-López (ORCID: https://orcid.org/0000-0003-0243-0816)
- Jennifer Angeline Gaddy (ORCID: https://orcid.org/0000-0002-2192-4224)
- Genesis Wilson (ORCID: https://orcid.org/0000-0001-7367-2324)
- Steven M. Damo (ORCID: https://orcid.org/0000-0002-1483-7712)
- Sandra Murray (ORCID: https://orcid.org/0000-0002-8267-9138)
- Celestine N. Wanjalla (ORCID: https://orcid.org/0000-0001-9159-5414)
- Blanca Rodríguez (ORCID: https://orcid.org/0000-0001-6361-3339)
- Elma Zaganjor (ORCID: https://orcid.org/0000-0003-0600-7417)
- Johnathan Moore (ORCID: https://orcid.org/0000-0002-3207-5360)
- Kit Neikirk (ORCID: https://orcid.org/0000-0003-0994-0333)
- Heather K. Beasley (ORCID: https://orcid.org/0000-0002-4636-2889)
- Margaret Mungai (ORCID: https://orcid.org/0000-0003-3292-1552)
- Zer Vue (ORCID: https://orcid.org/0000-0002-3502-8906)
- Desiree Nguyen
- Jianqiang Shao
- Amber Crabtree (ORCID: https://orcid.org/0000-0003-1156-1658)
- Jamaine Davis (ORCID: https://orcid.org/0000-0002-5078-1306)
- Bryanna Shao
- Lori Banks (ORCID: https://orcid.org/0000-0003-4684-8763)
- Serif Bacevac
- Dominique C. Stephens
- Kyrin Turner
- Chanel Harris
- Ashton Oliver (ORCID: https://orcid.org/0009-0001-9874-2621)
- Han Le
Institutions
- Meharry Medical College (US)
- University of Iowa (US)
- United States Department of Veterans Affairs (US)
- University of Pittsburgh (US)
- Vanderbilt University (US)
- Prairie View A&M University (US)
- Health First (US)
- VA Tennessee Valley Healthcare System (US)
- Vanderbilt University Medical Center (US)
- Fisk University (US)
Publication Details
- Journal
- Current Protocols
- Published
- 2025-02-01
- DOI
- https://doi.org/10.1002/cpz1.70004
- Citations
- 1
- Primary Topic
- Mitochondrial Function and Pathology
- Type
- article
- Field-Weighted Citation Impact
- 0.81
Funders
- National Science Foundation
- U.S. Department of Veterans Affairs
- Silicon Valley Community Foundation
- Bristol-Myers Squibb
- Vanderbilt University
- National Institutes of Health
- National Center for Research Resources
- National Center for Advancing Translational Sciences